Isolation and characterization of a membrane-attack-complex-inhibiting protein present in human serum and other biological fluids.
We have previously reported the isolation of a membrane-attack-complex-inhibiting protein (MIP) from human erythrocyte membranes [Watts, Patel & Morgan (1987) Complement 4, 236] and the production of polyclonal antibodies to this protein. Here we report the identification in plasma, urine, saliva and cerebrospinal fluid of a protein immunochemically identical with the membrane-derived MIP. The protein has been isolated from plasma by immunoaffinity chromatography on an anti-(erythrocyte MIP)-Sepharose column and shown by SDS/polyacrylamide-gel electrophoresis to be of similar molecular mass to the erythrocyte protein (55 kDa non-reduced and 65 kDa under reducing conditions). Monoclonal antibodies have been raised against plasma MIP and used to establish a two-site enzyme-linked immunoadsorbent assay, enabling quantification of MIP in plasma, urine and cerebrospinal fluid. Plasma MIP, though not able to incorporate spontaneously into membranes, was deposited on heterologous and homologous erythrocyte membranes during complement activation in a C8-dependent manner. Depletion of MIP from plasma resulted in enhancement of the lytic capacity of the plasma on heterologous erythrocytes.
Watts
M J
MJ
Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, U.K.
Dankert
J R
JR
Morgan
E P
EP
D000818:Animals; D000911:Antibodies, Monoclonal; D001798:Blood Proteins / Q000302:isolation & purification* / Q000652:urine; D018957:CD59 Antigens; D003185:Complement C8 / Q000302:isolation & purification; D003186:Complement C9 / Q000302:isolation & purification; D015938:Complement Membrane Attack Complex / Q000037:antagonists & inhibitors*; D003602:Cytotoxicity, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D004910:Erythrocyte Membrane / Q000276:immunology; D006801:Humans; D007700:Kinetics; D012463:Saliva / Q000276:immunology*; D012756:Sheep
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D018957:CD59 Antigens; D003185:Complement C8; D003186:Complement C9; D015938:Complement Membrane Attack Complex